DIHYDROXYACETONE PHOSPHATE (DHAP) IS AN IMPORTANT COMPOUND IN BIOCHEMISTRY AND ENZYMOLOGY. ENZYMATIC ALDOL REACTIONS HAVE DEMONSTRATED THAT DHAP IS REQUIRED AS A DONOR IN THE SYNTHESIS OF COMMON AND UNCOMMON SUGARS. GLYCEROL WAS FIRST CONVERTED TO GLYCEROL PHOSPHATE BY GLYCEROL KINASE. IN THIS REACTION, GLYCEROL KINASE AND CREATINE KINASE WERE IMMOBILIZED WITH A POLYACRYLIMIDE-CO-ACRYLOXYSUCCINIMIDE (PAN) CROSSLINKED GEL. THE REACTION TO PHOSPHORYLATE GLYCEROL WAS DONE ON A 1-7 MMOL SCALE AND IMMOBILIZED ENZYMES WERE USED, WITH CREATINE KINASE AND PHOSPHOCREATINE SERVING AS A REGENERATION SYSTEM FOR ATP. THIS REACTION DELIVERED THE PRODUCT, L-GLYCEROL PHOSPHATE, IN 100% CONVERSION. THIS PRODUCT WAS THEN CONVERTED TO DIHYDROXYACETONE PHOSPHATE IN A SUBSEQUENT ENZYMATIC REACTION. THE ENZYME FOR THIS PROCESS IS GLYCEROL PHOSPHATE OXIDASE, WHICH HAS A HIGH OXIDATION POTENTIAL AND, THEREFORE, FAVORED THE PRODUCTION OF DIHYDROXYACETONE PHOSPHATE. THE CONVERSION OF GLYCEROL PHOSPHATE TO DIHYDROXYACETONE PHOSPHATE WAS ACCOMPLISHED USING THIS OXIDASE IN A REACTION MIXTURE OF O2 SATURATED DISTILLED WATER, PH 6.8. GLYCEROL PHOSPHATE OXIDASE PRODUCES HYDROGEN PEROXIDE AS A BY PRODUCT, WHICH IS RECYCLED IN SITU TO OXYGEN AND HYDROGEN WITH CALASE. THIS REACTION YIELDED DIHYDROXYACETONE PHOSPHATE TO 100% CONVERSION, AND THE PRODUCT WAS ISOLATED BY BARIUM PRECIPTATION AND STORED AS THE BA SALT. THUS, UPON SUBSEQUENT REACTION, THIS PRODUCT WILL YIELD L-GLYCERALDEHYDE-3-PHOSPHATE IN ENANTIOMERIC PURITY. GENERATION OF THIS PRODUCT IS IN ITSELF IMPORTANT, AS IT IS A HIGHLY EXPENSIVE REAGENT, AND THUS HAS LIMITED USE IN SYNTHETIC REACTIONS. FURTHERMORE, TO THE BEST OF OUR KNOWLEDGE THIS REPRESENTS THE FIRST REPORTED SYNTHESIS OF 13C LABELED DIHYDROXYACETONE PHOSPHATE.